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phosphorylated smad3  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phosphorylated smad3
    Exogenous GDF11 induces muscle atrophy signaling in C2C12 myotubes. (A) GDF11 PPI network constructed using STRING. (B) GO-BP enrichment analysis derived from the STRING dataset. (C) GDF11 PPI network constructed using GeneMANIA. (D) GO-BP enrichment analysis derived from the GeneMANIA dataset. (E) Protein–protein docking simulation of the GDF11–ACVR2B complex. (F) Representative Western blot images of <t>p-SMAD3,</t> SMAD3, Atrogin-1, MuRF1, and Gapdh in C2C12 myotubes treated with rGDF11 (0–100 ng/mL) for 48 h. (G) Densitometric quantification of the p-SMAD3/SMAD3 protein ratio. (H) Densitometric quantification of Atrogin-1 protein expression. (I) Densitometric quantification of MuRF1 protein expression. (J,K) Relative mRNA expression of Atrogin-1 and MuRF1. Data are mean ± SEM. ns , not significant, * P < 0.05, ** P < 0.01, *** P < 0.001 versus control; n = 3.
    Phosphorylated Smad3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1609 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/phosphorylated+smad3/Phospho-Smad3+(Ser423%2F425)+Rabbit+mAb/pmc12975752-157-25-29
    Average 96 stars, based on 1609 article reviews
    phosphorylated smad3 - by Bioz Stars, 2026-10
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    Images

    1) Product Images from "Elevated circulating GDF11 and its role in age-related sarcopenia: insights from clinical, transcriptomic, and in vitro analyses"

    Article Title: Elevated circulating GDF11 and its role in age-related sarcopenia: insights from clinical, transcriptomic, and in vitro analyses

    Journal: Frontiers in Aging

    doi: 10.3389/fragi.2026.1736069

    Exogenous GDF11 induces muscle atrophy signaling in C2C12 myotubes. (A) GDF11 PPI network constructed using STRING. (B) GO-BP enrichment analysis derived from the STRING dataset. (C) GDF11 PPI network constructed using GeneMANIA. (D) GO-BP enrichment analysis derived from the GeneMANIA dataset. (E) Protein–protein docking simulation of the GDF11–ACVR2B complex. (F) Representative Western blot images of p-SMAD3, SMAD3, Atrogin-1, MuRF1, and Gapdh in C2C12 myotubes treated with rGDF11 (0–100 ng/mL) for 48 h. (G) Densitometric quantification of the p-SMAD3/SMAD3 protein ratio. (H) Densitometric quantification of Atrogin-1 protein expression. (I) Densitometric quantification of MuRF1 protein expression. (J,K) Relative mRNA expression of Atrogin-1 and MuRF1. Data are mean ± SEM. ns , not significant, * P < 0.05, ** P < 0.01, *** P < 0.001 versus control; n = 3.
    Figure Legend Snippet: Exogenous GDF11 induces muscle atrophy signaling in C2C12 myotubes. (A) GDF11 PPI network constructed using STRING. (B) GO-BP enrichment analysis derived from the STRING dataset. (C) GDF11 PPI network constructed using GeneMANIA. (D) GO-BP enrichment analysis derived from the GeneMANIA dataset. (E) Protein–protein docking simulation of the GDF11–ACVR2B complex. (F) Representative Western blot images of p-SMAD3, SMAD3, Atrogin-1, MuRF1, and Gapdh in C2C12 myotubes treated with rGDF11 (0–100 ng/mL) for 48 h. (G) Densitometric quantification of the p-SMAD3/SMAD3 protein ratio. (H) Densitometric quantification of Atrogin-1 protein expression. (I) Densitometric quantification of MuRF1 protein expression. (J,K) Relative mRNA expression of Atrogin-1 and MuRF1. Data are mean ± SEM. ns , not significant, * P < 0.05, ** P < 0.01, *** P < 0.001 versus control; n = 3.

    Techniques Used: Construct, Derivative Assay, Western Blot, Expressing, Control

    Related Articles

    SDS Page:

    Article Title: TMEPAI Confers Paclitaxel Resistance in Triple-Negative Breast Cancer Cells by Promoting AKT Phosphorylation and Its Downstream Cascade
    Article Snippet: .. Equal amounts of cell lysates were analyzed by SDS-PAGE with antibodies for β-actin, phosphorylated SMAD3, total SMAD3, phosphorylated AKT, total AKT, PI3K, Vimentin, E-Cadherin, and MRP-1 (Cell Signaling). .. Primary antibodies used were as follows: AKT (Cell Signaling Technology, CST #4685), phosphoAKT (Ser473) (CST #3787), β-Actin (CST #4967), GAPDH (CST #2118), caspase-3 (CST #9662), Cleaved caspase-3 (CST #9664), and TMEPAI (kindly provided by the Experimental Pathology Laboratory, University of Tsukuba).

    Article Title: FGF21 Ameliorates Fibroblasts Activation and Systemic Sclerosis by Inhibiting CK2α/GLI2 Signaling Axis.
    Article Snippet: Systemic sclerosis is a typical fibrotic disease of unknown etiology that is characterized by abnormal fibroblast activation and excessive deposition of extracellular matrix.. Unfortunately, effective therapeutic approaches are lacking.. FGF21 plays a key role in mediating a variety of biological activities.

    Control:

    Article Title: IFN-γ and TNF-α Impair Lung Development by Upregulating SMAD7 to Inhibit TGF-β Signaling Pathway and ECM Dysregulation
    Article Snippet: .. Primary antibodies were used as follows: α-smooth muscle actin (α-SMA, Cat#19245, 1:1000), collagen type I alpha 1 chain (COL1A1, Cat#91144, 1:1000), SMAD2 (Cat#5339, 1:1000), phosphorylated SMAD2 (p-SMAD2, Ser465/467, Cat#18338, 1:1000), SMAD3 (Cat#9523, 1:1000), phosphorylated SMAD3 (p-SMAD3, Ser423/425, Cat#9520, 1:1000), and SMAD4 (Cat#38454, 1:1000) from Cell Signaling Technology (CST, USA); SMAD7 (Cat#ab216428, 1:1000) from Abcam (UK); β-actin (1:1000) as loading control from Beijing Boaosen Biotechnology (Beigjing, China). .. Total RNA was extracted from cells/tissues with TRIzol reagent (Thermo Fisher Scientific) following the supplier’s instructions.

    Article Title: Elevated circulating GDF11 and its role in age-related sarcopenia: insights from clinical, transcriptomic, and in vitro analyses
    Article Snippet: .. Membranes were blocked with 5% skimmed milk for 1 h at room temperature and subsequently incubated with primary antibodies against Smad3 (ab40854, Abcam, United States), phosphorylated Smad3 (p-SMAD3; #9520, CST, United States), Atrogin-1 (67172-1-Ig, Proteintech, China), MuRF1 (55456-1-AP, Proteintech, China) and GAPDH (1E6D9, Proteintech, China), with GAPDH serving as the loading control. ..

    Article Title: IFN-γ and TNF-α Impair Lung Development by Upregulating SMAD7 to Inhibit TGF-β Signaling Pathway and ECM Dysregulation.
    Article Snippet: Following quality control, the prepared libraries were subjected to high-throughput sequencing on an Illumina Novaseq platform (Illumina, San Diego, CA, USA), generating 150 base-pair pairedend reads. .. For differential expression analysis, the edgeR 1 3 Page 3 of 15 75 Inflammation (2026) 49:75 SMAD3 (Cat#9523, 1:1000), phosphorylated SMAD3 (p-SMAD3, Ser423/425, Cat#9520, 1:1000), and SMAD4 (Cat#38454, 1:1000) from Cell Signaling Technology (CST, USA); SMAD7 (Cat#ab216428, 1:1000) from Abcam (UK); β-actin (1:1000) as loading control from Beijing Boaosen Biotechnology (Beigjing, China). .. Total RNA was extracted from cells/tissues with TRIzol reagent (Thermo Fisher Scientific) following the supplier’s instructions.

    Membrane:

    Article Title: FGF21 Ameliorates Fibroblasts Activation and Systemic Sclerosis by Inhibiting CK2α/GLI2 Signaling Axis.
    Article Snippet: Systemic sclerosis is a typical fibrotic disease of unknown etiology that is characterized by abnormal fibroblast activation and excessive deposition of extracellular matrix.. Unfortunately, effective therapeutic approaches are lacking.. FGF21 plays a key role in mediating a variety of biological activities.

    Western Blot:

    Article Title: FGF21 Ameliorates Fibroblasts Activation and Systemic Sclerosis by Inhibiting CK2α/GLI2 Signaling Axis.
    Article Snippet: Systemic sclerosis is a typical fibrotic disease of unknown etiology that is characterized by abnormal fibroblast activation and excessive deposition of extracellular matrix.. Unfortunately, effective therapeutic approaches are lacking.. FGF21 plays a key role in mediating a variety of biological activities.

    Article Title: UBE2O as a key regulator of drug-induced erythropoiesis in the context of myelodysplastic syndromes
    Article Snippet: .. Immunoblots were incubated overnight at 4°C with specific primary antibodies against vinculin (1:5000; catalog no. MA5-11690; Invitrogen), UBE2O (1:1000; catalog no. PA5-87583; Invitrogen), GATA1 (1:1000; catalog no. sc-266; Santa Cruz), β-actin (1:1000; catalog no. 4970S; Cell Signaling Technology), glyceraldehyde-3-phosphate dehydrogenase (1:1000; catalog no. sc-137179; Santa Cruz), and phosphorylated Smad3 (1:500; catalog no. 9520; Cell Signaling Technology) in Dulbecco’s PBS with 0.1% Tween 20, and protein detection was achieved using peroxidase-conjugated anti-mouse (catalog no. 7076S; Cell Signaling Technology), anti-rabbit (catalog no. 7074S; Cell Signaling Technology), or anti-rat (catalog no. 31470; Invitrogen) secondary antibodies, and a chemiluminescent reagent (catalog no. 170-5061; Bio-Rad). ..

    Article Title: UBE2O AS A KEY REGULATOR OF DRUG-INDUCED ERYTHROPOIESIS IN THE CONTEXT OF MYELODYSPLASTIC SYNDROMES.
    Article Snippet: .. Immunoblots were incubated overnight at 4°C with specific 130 primary antibodies against vinculin (1:5000, Invitrogen, #MA5-11690), UBE2O (1:1000, Invitrogen, #PA5-131 87583), GATA1 (1:1000, Santa Cruz, #sc-266), β-Actin (1:1000, Cell Signaling #4970S), GAPDH (1:1000, 132 Santa Cruz #sc-137179), and phosphorylated Smad3 (p-Smad3; 1:500, Cell Signaling Technology, #9520) in 133 Dulbecco’s PBS with 0.1% Tween 20, and protein detection was achieved using peroxidase-conjugated anti-134 mouse (Cell Signaling Technology, #7076S), anti-rabbit (Cell Signaling Technology, #7074S), or anti-rat 135 (Invitrogen, #31470) secondary antibodies and a chemiluminescent reagent (BioRad, #170–5061). ..

    Ubiquitin Proteomics:

    Article Title: FGF21 Ameliorates Fibroblasts Activation and Systemic Sclerosis by Inhibiting CK2α/GLI2 Signaling Axis.
    Article Snippet: Systemic sclerosis is a typical fibrotic disease of unknown etiology that is characterized by abnormal fibroblast activation and excessive deposition of extracellular matrix.. Unfortunately, effective therapeutic approaches are lacking.. FGF21 plays a key role in mediating a variety of biological activities.

    Incubation:

    Article Title: Elevated circulating GDF11 and its role in age-related sarcopenia: insights from clinical, transcriptomic, and in vitro analyses
    Article Snippet: .. Membranes were blocked with 5% skimmed milk for 1 h at room temperature and subsequently incubated with primary antibodies against Smad3 (ab40854, Abcam, United States), phosphorylated Smad3 (p-SMAD3; #9520, CST, United States), Atrogin-1 (67172-1-Ig, Proteintech, China), MuRF1 (55456-1-AP, Proteintech, China) and GAPDH (1E6D9, Proteintech, China), with GAPDH serving as the loading control. ..

    Article Title: UBE2O as a key regulator of drug-induced erythropoiesis in the context of myelodysplastic syndromes
    Article Snippet: .. Immunoblots were incubated overnight at 4°C with specific primary antibodies against vinculin (1:5000; catalog no. MA5-11690; Invitrogen), UBE2O (1:1000; catalog no. PA5-87583; Invitrogen), GATA1 (1:1000; catalog no. sc-266; Santa Cruz), β-actin (1:1000; catalog no. 4970S; Cell Signaling Technology), glyceraldehyde-3-phosphate dehydrogenase (1:1000; catalog no. sc-137179; Santa Cruz), and phosphorylated Smad3 (1:500; catalog no. 9520; Cell Signaling Technology) in Dulbecco’s PBS with 0.1% Tween 20, and protein detection was achieved using peroxidase-conjugated anti-mouse (catalog no. 7076S; Cell Signaling Technology), anti-rabbit (catalog no. 7074S; Cell Signaling Technology), or anti-rat (catalog no. 31470; Invitrogen) secondary antibodies, and a chemiluminescent reagent (catalog no. 170-5061; Bio-Rad). ..

    Article Title: UBE2O AS A KEY REGULATOR OF DRUG-INDUCED ERYTHROPOIESIS IN THE CONTEXT OF MYELODYSPLASTIC SYNDROMES.
    Article Snippet: .. Immunoblots were incubated overnight at 4°C with specific 130 primary antibodies against vinculin (1:5000, Invitrogen, #MA5-11690), UBE2O (1:1000, Invitrogen, #PA5-131 87583), GATA1 (1:1000, Santa Cruz, #sc-266), β-Actin (1:1000, Cell Signaling #4970S), GAPDH (1:1000, 132 Santa Cruz #sc-137179), and phosphorylated Smad3 (p-Smad3; 1:500, Cell Signaling Technology, #9520) in 133 Dulbecco’s PBS with 0.1% Tween 20, and protein detection was achieved using peroxidase-conjugated anti-134 mouse (Cell Signaling Technology, #7076S), anti-rabbit (Cell Signaling Technology, #7074S), or anti-rat 135 (Invitrogen, #31470) secondary antibodies and a chemiluminescent reagent (BioRad, #170–5061). ..

    Quantitative Proteomics:

    Article Title: IFN-γ and TNF-α Impair Lung Development by Upregulating SMAD7 to Inhibit TGF-β Signaling Pathway and ECM Dysregulation.
    Article Snippet: Following quality control, the prepared libraries were subjected to high-throughput sequencing on an Illumina Novaseq platform (Illumina, San Diego, CA, USA), generating 150 base-pair pairedend reads. .. For differential expression analysis, the edgeR 1 3 Page 3 of 15 75 Inflammation (2026) 49:75 SMAD3 (Cat#9523, 1:1000), phosphorylated SMAD3 (p-SMAD3, Ser423/425, Cat#9520, 1:1000), and SMAD4 (Cat#38454, 1:1000) from Cell Signaling Technology (CST, USA); SMAD7 (Cat#ab216428, 1:1000) from Abcam (UK); β-actin (1:1000) as loading control from Beijing Boaosen Biotechnology (Beigjing, China). .. Total RNA was extracted from cells/tissues with TRIzol reagent (Thermo Fisher Scientific) following the supplier’s instructions.



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    Molecular modeling and pathological consequences of the TGFBR1 p.Asp351Tyr variant. (A) Predicted structural models of the interaction between wild‐type (WT) TGFBR1 and its substrates <t>SMAD2</t> (left) and <t>SMAD3</t> (right). Insets provide detailed views of the key interacting residues at the binding interface. (B) Predicted structural models of the p.Asp351Tyr (D351Y) mutant TGFBR1 with SMAD2 (top) and SMAD3 (bottom). The mutation is predicted to significantly alter the binding interface and increase interactions with SMAD2, whereas the interaction with SMAD3 remains largely unchanged. (C) Histopathological analysis of the patient′s aortic wall compared to a control aorta. Hematoxylin and eosin (HE) staining reveals disorganization of the medial layer. Elastica van Gieson (EvG) staining shows severe fragmentation and loss of elastic fibers. Masson′s trichrome staining indicates increased collagen deposition (blue). Scale bar = 200 μ m. (D) Immunofluorescence staining for <t>phosphorylated</t> <t>SMAD2/SMAD3</t> (p‐SMAD2/SMAD3, red), a marker of TGF‐ β pathway activation. Compared to the control, the patient′s aortic tissue shows a marked increase in nuclear p‐SMAD2/SMAD3 signal (indicated by white arrowheads) in smooth muscle cells ( ACTA2 , green). Nuclei are counterstained with DAPI (blue). Scale bar = 100 μ m.
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    Image Search Results


    Molecular modeling and pathological consequences of the TGFBR1 p.Asp351Tyr variant. (A) Predicted structural models of the interaction between wild‐type (WT) TGFBR1 and its substrates SMAD2 (left) and SMAD3 (right). Insets provide detailed views of the key interacting residues at the binding interface. (B) Predicted structural models of the p.Asp351Tyr (D351Y) mutant TGFBR1 with SMAD2 (top) and SMAD3 (bottom). The mutation is predicted to significantly alter the binding interface and increase interactions with SMAD2, whereas the interaction with SMAD3 remains largely unchanged. (C) Histopathological analysis of the patient′s aortic wall compared to a control aorta. Hematoxylin and eosin (HE) staining reveals disorganization of the medial layer. Elastica van Gieson (EvG) staining shows severe fragmentation and loss of elastic fibers. Masson′s trichrome staining indicates increased collagen deposition (blue). Scale bar = 200 μ m. (D) Immunofluorescence staining for phosphorylated SMAD2/SMAD3 (p‐SMAD2/SMAD3, red), a marker of TGF‐ β pathway activation. Compared to the control, the patient′s aortic tissue shows a marked increase in nuclear p‐SMAD2/SMAD3 signal (indicated by white arrowheads) in smooth muscle cells ( ACTA2 , green). Nuclei are counterstained with DAPI (blue). Scale bar = 100 μ m.

    Journal: Human Mutation

    Article Title: Type B Aortic Dissection Following Abdominal Aortic Aneurysm Repair in Loeys–Dietz Syndrome: A Novel TGFBR1 Variant

    doi: 10.1155/humu/2951304

    Figure Lengend Snippet: Molecular modeling and pathological consequences of the TGFBR1 p.Asp351Tyr variant. (A) Predicted structural models of the interaction between wild‐type (WT) TGFBR1 and its substrates SMAD2 (left) and SMAD3 (right). Insets provide detailed views of the key interacting residues at the binding interface. (B) Predicted structural models of the p.Asp351Tyr (D351Y) mutant TGFBR1 with SMAD2 (top) and SMAD3 (bottom). The mutation is predicted to significantly alter the binding interface and increase interactions with SMAD2, whereas the interaction with SMAD3 remains largely unchanged. (C) Histopathological analysis of the patient′s aortic wall compared to a control aorta. Hematoxylin and eosin (HE) staining reveals disorganization of the medial layer. Elastica van Gieson (EvG) staining shows severe fragmentation and loss of elastic fibers. Masson′s trichrome staining indicates increased collagen deposition (blue). Scale bar = 200 μ m. (D) Immunofluorescence staining for phosphorylated SMAD2/SMAD3 (p‐SMAD2/SMAD3, red), a marker of TGF‐ β pathway activation. Compared to the control, the patient′s aortic tissue shows a marked increase in nuclear p‐SMAD2/SMAD3 signal (indicated by white arrowheads) in smooth muscle cells ( ACTA2 , green). Nuclei are counterstained with DAPI (blue). Scale bar = 100 μ m.

    Article Snippet: After blocking, they were incubated overnight at 4°C with a primary antibody against phosphorylated SMAD2/SMAD3 (ABclonal, AP0548).

    Techniques: Variant Assay, Binding Assay, Mutagenesis, Control, Staining, Immunofluorescence, Marker, Activation Assay

    Exogenous GDF11 induces muscle atrophy signaling in C2C12 myotubes. (A) GDF11 PPI network constructed using STRING. (B) GO-BP enrichment analysis derived from the STRING dataset. (C) GDF11 PPI network constructed using GeneMANIA. (D) GO-BP enrichment analysis derived from the GeneMANIA dataset. (E) Protein–protein docking simulation of the GDF11–ACVR2B complex. (F) Representative Western blot images of p-SMAD3, SMAD3, Atrogin-1, MuRF1, and Gapdh in C2C12 myotubes treated with rGDF11 (0–100 ng/mL) for 48 h. (G) Densitometric quantification of the p-SMAD3/SMAD3 protein ratio. (H) Densitometric quantification of Atrogin-1 protein expression. (I) Densitometric quantification of MuRF1 protein expression. (J,K) Relative mRNA expression of Atrogin-1 and MuRF1. Data are mean ± SEM. ns , not significant, * P < 0.05, ** P < 0.01, *** P < 0.001 versus control; n = 3.

    Journal: Frontiers in Aging

    Article Title: Elevated circulating GDF11 and its role in age-related sarcopenia: insights from clinical, transcriptomic, and in vitro analyses

    doi: 10.3389/fragi.2026.1736069

    Figure Lengend Snippet: Exogenous GDF11 induces muscle atrophy signaling in C2C12 myotubes. (A) GDF11 PPI network constructed using STRING. (B) GO-BP enrichment analysis derived from the STRING dataset. (C) GDF11 PPI network constructed using GeneMANIA. (D) GO-BP enrichment analysis derived from the GeneMANIA dataset. (E) Protein–protein docking simulation of the GDF11–ACVR2B complex. (F) Representative Western blot images of p-SMAD3, SMAD3, Atrogin-1, MuRF1, and Gapdh in C2C12 myotubes treated with rGDF11 (0–100 ng/mL) for 48 h. (G) Densitometric quantification of the p-SMAD3/SMAD3 protein ratio. (H) Densitometric quantification of Atrogin-1 protein expression. (I) Densitometric quantification of MuRF1 protein expression. (J,K) Relative mRNA expression of Atrogin-1 and MuRF1. Data are mean ± SEM. ns , not significant, * P < 0.05, ** P < 0.01, *** P < 0.001 versus control; n = 3.

    Article Snippet: Membranes were blocked with 5% skimmed milk for 1 h at room temperature and subsequently incubated with primary antibodies against Smad3 (ab40854, Abcam, United States), phosphorylated Smad3 (p-SMAD3; #9520, CST, United States), Atrogin-1 (67172-1-Ig, Proteintech, China), MuRF1 (55456-1-AP, Proteintech, China) and GAPDH (1E6D9, Proteintech, China), with GAPDH serving as the loading control.

    Techniques: Construct, Derivative Assay, Western Blot, Expressing, Control